Fig. 6
Intracellular localization of Snx3 mutants, and their effects on EGF receptor degradation. a Cells were transiently transfected with cDNAs coding for the indicated GFP-SNX3 fusion proteins. 18 h after transfection cells were starved for 4 h, challenged with EGF for 15 or 120 min, fixed and stained with anti-EGF recetptor antibodies. The cells were then analyzed by automated microscopy. Bar: 10 µm. b, c In a, the total GFP fluoresence intensity of endosomes containing the EGF receptor after 15 min was quantified per cell (b) and is compared to the total intensity of the GFP fluorescence signal per cell (c). d The experiments was as in a for the indicated time periods. The total EGF receptor per cell was quantified in transfected cells (solid line) and in neighboring untransfected cells (dashed line) from the same well of the 96-well plates. The integrated intensity is expressed as a percentage of the values observed at 15 min. b–d Aproximately 1000 cells were analysed per condition in n = 3 independent experiments. Error bars indicate SEM. Statistical significance is calculated using one-way ANOVA analysis with Bonferonni’s post-test. Levels of significance are indicated as follows: *p < 0.05; ***p < 0.001
